Use of Histones for Therapeutic Purposes

ABSTRACT

The invention relates to the use of at least one human recombinant histone, especially at least one histone H1 subtype, and/or a therapeutic histone fraction as a basis for the treatment of thrombocytopenia.

The present application is a continuation of U.S. patent applicationSer. No. 13/758,950 filed Feb. 4, 2013, which is a continuation of U.S.patent application Ser. No. 11/920,050; filed Nov. 6, 2008 nowabandoned, which is a national stage entry of PCT/EP2006/004167; filedMay 4, 2006, which claims priority to DE 10 2005 022 319.2; filed May10, 2005, all which are hereby incorporated by reference.

The invention relates to the use of at least one human recombinanthistone of H1 subtype and/or of its therapeutically effective segment,especially histone H1.3 for therapeutic purposes.

The use of therapeutic active substances based on human recombinanthistone H1 subtypes for the treatment of cancers, e.g. leukemia, isdisclosed in the article by Reiner Class et al. in Am. J. Clin. Oncol.(CCT) vol. 19 No. 5 1996 and European patent application 98919254.7.

The effect of histone H1 and H2A/H2B fractions from calf thymus onhematopoietical stem cells (CFU-S) in normal and radioactivelyirradiated rats was described in a Russian article by Semina 0. V. etal. in Radiatsionnaia Biologiia, Radioecologiia 34 (4-5), 1994,July-October.

In complex pathological conditions such as acute myeloid leukemia, thereis frequently observed to be a thrombocytopenia which may even beenhanced by a chemotherapeutic treatment of the leukemia.Thrombocytopenia is, however, also observed with other etiology. Sincethrombocytopenia may lead to life-threatening internal hemorrhages,therapies for various pathological symptoms which may be the cause ofthe thrombocytopenia are frequently made particularly difficult.

It was therefore an object of the invention to find an active substancewhich can be used therapeutically for thrombocytopenia in order thusalso to improve substantially the success of curing the basic symptom asinducer of the thrombocytopenia.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 (hereinafter “FIG. 1”) shows the effect of the active substancesof the invention in a 0.9% NaCl solution at, 37.5 mg/qm² of body surfacearea on the thrombocytopenia of a patient.

DETAILED DESCRIPTION

It has been possible according to the invention to achieve the object byusing for the therapy of thrombocytopenia inter alia as a result offaulty stem cell differentiation or weakened proliferation ofmegakaryocytes an active substance based on at least one humanrecombinant histone (especially at least one histone of H1 subtype)and/or its therapeutically effective segment.

This applies especially to a thrombocytopenia as concomitantmanifestation of a hematological disorder.

It is moreover possible to employ the therapeutic method of theinvention for the treatment of thrombocytopenia during or afterchemotherapy for the treatment of a hematological disorder, especiallyacute myeloid leukemia.

It has surprisingly been possible to show that the active substance ofthe invention shows on the one hand a positive result in the treatmentof a hematological disorder such as leukemia, but on the other hand alsoa positive result in the treatment of the thrombocytopenia associatedwith the hematological disorders.

It was thus possible with one and the same active substance to note botha regression in the leukemia and an increase in platelets.

It was possible to achieve therapeutic trial results also on patientswith human recombinant histone H1.3, it having been possible to reducemarkedly the number of pathological tumor cells in an AML patient and,at the same time, to increase substantially platelet production, wherebyit was possible to improve substantially the prospects of curing thepatient.

The trial results are reproduced in more detail below, the activesubstance used in this case, based on human recombinant H1.3concentrations 37.5 mg/qm² of body surface area etc., having beenadministered in a 0.9% NaCl solution intravenously 3 times a week over aperiod of about 4 hours.

FIG. 1 shows the effect of the active substances of the invention in a0.9% NaCl solution at, 37.5 mg/qm² of body surface area on thethrombocytopenia of a patient.

The platelets in peripheral blood are on the ordinate in a number offrom 0 to 40×10⁹. Treatment with active substance of the invention forthree weeks is shown on the abscissa, the platelet count measured forthe patient before the first treatment being greatly reduced at about8×10⁹. Then in each case three drip infusions take place per week within each case 4 hours per infusion, with the 1st to 3rd infusion on the1st, 3rd and 5th day in the first week, with the 4th to 6th infusion onthe 8th, 10th and 12th day in the second week and finally with the 7thto 9th infusion on the 15th, 17th and 19th day in the 3rd week. On the29th day after the first infusion, a control measurement of the plateletcount took place without a further infusion with the active substance ofthe invention. At a later time FU1, the patient was discharged with analmost normal platelet count of about 34×10⁹, without an activesubstance of the invention having been supplied even once. This valuelay outside the need to supply stored blood. The patient was asked toattend a follow-up examination with the possibility of resumption oftreatment if the platelet count has not improved further on its own orhad even deteriorated. The results of the follow-up examination are notshown here.

The appended FIG. 1 shows at the start of the second week up toconclusion of the treatment in the third week a jump in the plateletcount after the 5th day of treatment and then a continuous rise in theplatelet count from the 8th to the subsequent 19th day of treatment anda further slower rise in the platelet count at the first controlexamination on the 29th day and at a later discharge day FU1 of thepatient without further addition of the active substance of theinvention, the finally measured platelet count being, as already stated,about 32.5×10⁹.

The invention is not restricted to the use of human recombinant H1.3.Because of the close relationship of the H1 subtypes, it is obvious to askilled worker also to employ as active substance other humanrecombinant H1 subtypes as basis for the active substance of theinvention.

Following the successful therapy of patients with a thrombocytopenia,here as concomitant syndrome of an AML leukemia, with human recombinanthistone H1.3, it is particularly obvious to a skilled worker also toemploy other recombinant H1 subtypes as alternative active substancessingly or in combination according to the invention.

The active substance of the invention preferably consists of thecomplete unshortened subtypes of histone proteins. However, it is alsoobvious to a skilled worker to look for the therapeutically effectivesegment, of which he is capable directly on the basis of his expertknowledge and experience without outstanding innovative contributionsbeing necessary in this case. Such therapeutically effective histonesegments therefore lie within the range of equivalents of the teachingof the invention disclosed herein.

The invention further discloses the therapeutic teaching of employing,when there is a threatening or incipient primary disorder which,experience has shown, may result in thrombocytopenia, the activesubstance of the invention prophylactically against a threateningthrombocytopenia even if, unlike leukemia, the active substance of theinvention is not effective against the primary disorder.

1-7. (canceled)
 8. A method for treating thrombocytopenia in a patientin need thereof, the method comprising: administering to the patient atleast three doses of a pharmaceutical composition comprising histoneH1.3 or a therapeutically active histone H1.3 segment during a period ofat least 8 days, wherein administration of the pharmaceuticalcomposition increases the number of platelets in the peripheral blood,thereby treating the thrombocytopenia.
 9. The method according to claim8, wherein the thrombocytopenia results from a hematological disorder.10. The method according to claim 9, wherein the hematological disorderis a leukemia.
 11. The method according to claim 10, wherein theleukemia is an acute myeloid leukemia.
 12. The method according to claim8, wherein the treatment of the thrombocytopenia occurs concurrentlywith or after a therapy for a hematological disorder.
 13. The methodaccording to claim 12, wherein the therapy is a chemotherapy.
 14. Themethod according to claim 12, wherein the hematological disorder is aleukemia.
 15. The method according to claim 14, wherein the leukemia isan acute myeloid leukemia.
 16. The method according to claim 8, whereinthe administration is intravenous.
 17. The method according to claim 8,further comprising the administration of other human recombinantsubtypes of histone H1 in combination with the histone H1.3 ortherapeutically active histone H1.3 segment.
 18. A method for treatingthrombocytopenia in a patient in need thereof caused by a primarydisorder, comprising: administering to the patient a pharmaceuticalcomposition comprising histone H1.3 or a therapeutically active histoneH1.3 segment, wherein the histone H1.3 is not effective to treat theprimary disorder.
 19. The method according to claim 18, wherein thethrombocytopenia results from a hematological disorder.
 20. The methodaccording to claim 18, wherein the treatment of the thrombocytopeniaoccurs concurrently with or after a therapy for a hematologicaldisorder.
 21. The method according to claim 20, wherein the therapy is achemotherapy.
 22. The method according to claim 18, whereinadministration to the patient is administration of at least three dosesof a pharmaceutical composition comprising histone H1.3 or atherapeutically active histone H1.3 segment during a period of at least8 days.
 23. The method according to claim 18, wherein the administrationis intravenous.
 24. The method according to claim 18, further comprisingthe administration of other human recombinant subtypes of histone H1 incombination with the histone H1.3 or therapeutically active histone H1.3segment.
 25. A method for treating threatening or incipientthrombocytopenia in a patient which is expected to be caused bychemotherapy, comprising: administering to the patient a pharmaceuticalcomposition comprising histone H1.3 or a therapeutically active histoneH1.3 segment concurrently with chemotherapy.
 26. The method according toclaim 25, wherein the administration is intravenous.
 27. The methodaccording to claim 25, further comprising the administration of otherhuman recombinant subtypes of histone H1 in combination with the histoneH1.3 or therapeutically active histone H1.3 segment.